Respiratory Pigments in Animals: Relation Structure-Function

185 Pages·1985·26.424 MB·English

by  W. A. Hendrickson, J. L. Smith, S. Sheriff (auth.), Professor Dr. Jean Lamy, Professor Dr. Jean-Paul Truchot, Professor Dr. Raymond Gilles (eds.)

IUBS Section of Comparative Pbysiology and Biochemistry
lst International Congress, Liege, Belgium, August 27-31, 1984
Conference OrgaDization
Organizing Board
R. Gilles, Chairman, Liege, Belgium
M. Gilles-Baillien and L. Bolis, Liege, Belgium/Messina, Italy
Host Society
European Society for Comparative Physiology
and Biochemistry
Under the Patronage of
The European Economic Community
The Fonds National de la Recherche Scientifique
The Ministere de l'Education Nationale et de la Culture Franyaise
The Fondation Uon Fredericq
The University of Liege
The European Society for Comparative Physiology and Biochemistry
The American Society of Zoologists
The Canadian.Society of Zoologists
The Japanese Society for General and Comparative Physiology
The Congress has been organized in relation with the tOOth Anniversary
of the School of Comparative Physiology and Biochemistry of the
University of Liege.
The proceedings of the invited lectures to the different symposin of the
congress have been gathered in five different volumes published by
Springer-Verlag under the following titles:
Circulation, Respiration, and Metabolism
Current Comparative Approaches
Edited by R. Gilles (ISBN 3-540-15627-5)
Transport Processes, 10nOo and Osmoregulation
Current Comparative Approaches
Edited by R. Gilles and M. Gilles-Baillien (ISBN 3-540-15628-3)
Neurobiology, Current Comparative Approaches
Edited by R. Gilles and J. Balthazart (ISBN 3-540-15480-9)
Respiratory Pigments in Animals, Relation Structure-Function
Edited by J. Lamy, J.-P. Truchot, and R. Gilles (ISBN 3-540-15629-1)
Higb Pressure Effects on Selected Biological Systems
Edited by A.J. R. Pequeux and R. Gilles (ISBN 3-540-15630-5)
Respiratory Pigments
in Animals
Relation Structure-Function
Edited by
J. Lamy, J.-P. Truchot, and R. Gilles
With 63 Figures
Springer-Verlag
Berlin Heidelberg New York Tokyo
Professor Dr. JEAN LAMY (Scientific Editors)
Laboratory of Biochemistry
Department of Pharmaceutical Science
University of Francois-Rabelais
2, Boulevard Tonnelle
F-37032 Tours Cedex, France
Professor Dr. JEAN-PAUL TRUCHOT
Laboratory of Neurobiology and Compared Physiology
University of Bordeaux I
Place du Dr. Bertrand Peyneau
F-33120 Arcachon Cedex, France
Professor Dr. RAYMOND GILLES (Coordinating Editor)
Laboratory of Biochemistry
University of Liege
22, Quai Van Benden
B-4020 Liege, Belgium
ISBN-13: 978-3-540-15629-1 e-ISBN-13: 978-3-642-70616-5
001: 10.1007/978-3·642-70616-5
Library of Congress Cataloging-in-PubLication Dala. Main entry under title: Respiratory
pigments in anUnab. Partial proceedings of the First International Congress of Comparative
Physiology and Biochemistry organized at Li~ge. Belgium, in August 1984 under the auspices
of the Section 01 Comparative Physiology and Biochemistry of the International Union of
Biological Sciences. Includes Index. I. Blood-Pigments-Structure_activity relationships
Congresses_ 2.Hemocyanin-StruclUre-activiry relationships-Congresses. 3. Hemoglobin
Sttucture-activiry relationships-Congresses. I. Lamy, Jean, 1941-. II. Truchot, Jean-Paul,
1937-. m. Gi1Jes, R.IV.lntemational Congress of Comparative Physiology and Bioehemimy
(lsi: 1984 : Litge, Belgium) V. International Union of Biological Sciences. Section of
Comparative Pbysiology and Biochemistry. QP99.3.P5R471935 591.1'13 85--22075
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CI by Springer-Verlag Berlin Heidelberg 1935
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Foreword
This volume is one of those published from the proceedings of the invited lectures to
the First International Congress of Comparative Physiology and Biochemistry I organized
at LI~e (Belgium) in August 1984 under the auspices of the Section of Comparative
Physiology and Biochemistry of the International Union of Biological Sciences. In a
general foreword to these different volumes, it seems to me appropriate to consider briefly
what may be the comparative approach.
Living organisms, beyond the diversity of their morphological forms, have evolved a
widespread range of basic solutions to cope with the different problems, both organisma!
and environmental, with which they are faced. Soon after the turn of the century, some
biologists realized that these solutions can be best comprehended In the framework of a
comparative approach integrating results of phYSiological and biochemical studies done at
the organismic, cellular and molecular levels. The development of this approach amongst
both physiologists and biochemists remained, however, extremely slow until recently.
Physiology and biochemistry have indeed long been mainly devoted to the service of med·
icine, finding scope enough for their activities in the study of a few species, particularly
mammals. This has tended to keep many physiologists and biochemists from the com·
parative approach, which demands either the widest possible survey of animal forms or
an integrated knowledge of the specific adaptive features of the specIes considered. These
particular characteristics of the comparative approach have, on the other hand, been very
attractive for biologists interested in the mechanisms of evolution and environmental
adaptations. This diversity of requirements of the comparative approach, at the conceptual
as well as at the technological level, can easily account for the fact that it emerged only
slowly amongst the other new, more rapidly growing, disciplines of the biological sciences.
Although a few pioneers have been working in the field since the beginning of the century,
it only started effectively in the early 1960's. 1960 was the date of the organization of the
periodical "Comparative Physiology and Biochemistry" by Kerkut and Scheer and of the
publication of the first volumes of the comprehensive treatise "Comparative Biochemistry"
edited by Florkin and Mason. These publications can be considered as milestones in the
evolution of the comparative approach. They have been followed by many others which
have greaUy contributed to giving the field the international status it deserved. Since the
1960's, the comparative approach has been maturing and developing more and more
VI
rapidly Into the Independent discipline it now is, widely recognized by the international
communities of physiologists, biochemists and biologists. It is currently used as an effective
tool of great help In the understanding of many research problems: biological as well as
clinical, applied as well illS fundamental.
The actual development of the field and the interest it arouses in ill growing portion of
the biological scientific community led some of us to consider the organization of an inter·
national structure, bringing together the major representative societies and groups around
the world, which would aim at the general advancement ilInd promotion of the comparative
approach. This was done in 1979 with the incorporation, within the international Union of
Biological Sciences, of a Section of Comparative Physiology and Biochemistry. The first
International Congress of CPB, I organized in Li~e with the help of a few friends and col
leagues, is the first activity of this newly founded Section. In 22 symposia it gathered some
146 invited lectures given by internationally renowned scientists on all major current topics
and trends in the field. The proceedings of these lectures have been collected in 5 volumes
produced by Springer-Verlag, a publisher long associated with the development of CPB.
The organization of the CPB Section of IUBS, Its first Congress and these proceedings
volumes can well be considered as milestones reflecting the international status and the
maturity that the comparative approach has gained, as a recognized independent discipline,
in the beginning of the 1980's, some 20 years after It was effectively launched.
Finally, I would like to consider that the selection of Llilge for this first Intemational
Congress has not been simply coincidental. I thus feel that this brief foreword would not
be complete without noting the privileged role U~e has played In some events associated
with the development of the comparative approach. LlAge had a pioneer in comparative
physiology already at the end of the last century with Leon Fredericq. With Marcel Florkin,
LlAge had its first Professor of biochemistry and one of the founding fathers of comparative
biochemistry. These two major figureheads of the comparative approach founded and devel
oped what is actually called the LlAge School of Comparative Physiology and Biochemistry,
which was, at the time of the Congress, celebrating its lOOth anniversary. This school provided
early support to the European Society for Comparative Physiology and Biochemistry orga
nized by Marcel Florkln and myself some years ago. The soclety •. stlll headquartered In LI~e,
was, with the CPBdlvlslon of the American Society of Zoologists, at the origin of the forma
tion of the CPB Section of I U BS under the auspices of which this first I nternational Congress,
specifically devoted to the comparative approach, has been organized. An essential particu·
larly of the LlAge school of CPB is that its two founding father~! SCientists interested In
general, basic aspects of the organization of living organisms, were also professors at the
faculty of medicine. This largely contributed in LlAge to avoiding the undesirable structura
tion of a 5O<alled "zoophyslology" or "zoobiochemistry" independent of the rest of the
field. The conditions were thus realized very early In LiAge for CPB to play its key role in
canalizing the necessary interactions between the general, pre-clinical or clinical and the
environmental, ecological or evolutionary tendencies of physiology and biochemistry. The
possibility of stimulating such interactions has served as a major guide line in the selection
of the symposia and invited lectures from which these proceedings have issued.
August, 1985 R. GILLES
Preface
Evolution has exploited the coordination chemistry of transition metals in many
subtle ways. Some of the most ingenious and original solutions of chemical problems are
to be found in the designs of metal complexes which combine reversibly with molecular
oxygen. Supposing an inorganic chemist ignorant of biochemistry had been asked which
metal Nature was most likely to have used in making an oxygen-carrylng protein, he would
probably have excluded iron from his list of guesses In view of its liability to become irre
versibly oxidised. Yet this Is precisely the metal that Nature has used in nearly all species
perhaps because of the ease of its change from five to six coordination and the facility with
which the strength of its bond with oxygen can be tuned to physiological needs.
Combination of ferrous iron with molecular oxygen needs donation of electron den
sity from the iron to the oxygen so that the complex acquires partial Fe+·Ol character.
Coordination of the iron to the free nitrogen of a porphyrin and to the imidazole of a
histidine has provided just the right degree of electron donation for an iron oxygen bond
strength of about one fifth of that of a carbon-carbon single bond. For optimum strength
the Fe-Q-Q angle should be about 120°. The partial negative charge on the oxygen allows
it to form hydrogen bonds with donor groups such as histidines or glutamines. The change
of the iron from five to six coordination is accompanied by a shortening of the bonds
between the iron and the porphyrin nitrogen5 and a movement of the iron relative to the
plane of the porphyrin nitrogens, from out of plane in the deoxygenated to in plane in the
oxygenated state.
These properties have offered Nature several ways of tuning the oxygen affinity. It
can tailor the distal heme pocket so as to vary the steric hindrance to the binding of
oxygen, it can alter the number and strength of hydrogen bonds to the bound oxygen and
it can design the protein so as to promote or hinder the movement of the iron into the
porphyrin plane. The combination of these devices has resulted in a range of oxygen
equilibrium constants that vary over at least four orders of magnitude from Ascaris hemo
globin with Ka = 145 to human hemoglobin in the T structure with Ka e 0.008. The
equilibrium constant of trout IV homoglobin at acid pH may be several orders smaller still.
Hemerythrins have evolved an entirely different coordination complex, consisting of
a pair of iron atoms bridged by two carboxylate ions and an oxygen atom, and coordinated
to five histidines of the protein, leaving one coordination site at one of the irons open for
VIII
coordination with oxygen. It seems that this iron-protein complex combines with oxygen
without undergoing a significant change of structure.
The pair of copper atoms that form the active site of hemocyanin probably form a
bridged dioxygen complex that links the two copper atoms at a distance of :! 3.5 .2..
EXAFS shows that dissociation of oxygen is accompanied by a stereochemical change,
probably consisting of a movement apart from the two copper atoms. The precise coordi
nation of the copper atoms is not yet known. The third metal used for reversible combina
tion with oxygen is vanadium, but the chemistry of oxygen carriers containing this metal
is still unexplored.
Once evolution had solved the problem of designing transition metal complexes capa
ble of reversible combination with oxygen, the next step essential for the evolution of fast
moving animals was the development of cooperative oxygen binding. One could well imagine
an oxygen carrier made up of hemes in parallel pockets of a single polypeptide, joined and
geared so that movement of anyone heme iron towards the porphyrin plane pushes all the
others in the same direction and thus raises their oxygen affinity. Why has Nature never
adopted such a design? Perhaps because such a simple sequential mechanism does not lend
itself to the generation of heterotropic cooperative effects needed for regulation of the
oxygen affinity and evolution of two-way respiratory carriers.
I n every instance discovered so far, cooperative effects are exhibited only by oligomeric
proteins in equilibrium between alternative states with different oxygen affinities; these
may be either alternative quaternary structures or alternative states of aggregation. Such
alternative states arise only if combination of the metal with oxygen is accompanied by a
stereochemical change at the metal or at the protein surrounding it. Consequently coopera
tive effects are exhibited only by hemoglobin and hemocyanins but not by hemerythrins.
A further important step in evolution has been the enclosure of oxygen carriers in
erythrocytes. In order to contain oxygen carriers in the blood vessels they had to have very
large molecular weights. Judging by the properties of erythrocruorins and hemocyanins,
such large complexes do not exhibit as much chemical versatility as the 0: tj tetramer In
erythrocytes.
The pages in this volume show how fantastically adaptable hemoglobin compared to
hemocyanin and hemerythrin. Not only does its oxygen affinity vary over an enormous
range, but the number of subunits per molecule and with them the free energy of coopera
\lvity, the strength and direction of the Bohr effect, the nature and binding energy of the
allosteric effectors and even the thermodynamics of the reactiqn with oxygen vary tre
mendously. Perhaps this is why animals with hemoglobin have evolved so successfully.
One of the most impressive features of the hemoglobins is the constancy of their
tertiary structure, ranging all the way from leg-hemoglobins that are coded for by plant
genes, to Invertebrate and vertebrate hemoglobins. One would have imagined this con
stancy to be· the result of an invariant amino acid sequence, but in fact the only invariant
residues are the proximal histidine and a phenylalanine that wedges the heme Into its
pocket. The only feature common to aU the hemoglobins is a set of internal sites which are
invariably occupied by non-polar amino acid residues ensuring the exctusion of polar
residues from the interior. This appears to have been sufficient to preserve the vitally
IX
important geometry of the heme pocket, but allowed the structures to vary in detail. The
angles between helical segments differ by up to 200 and the points of contact between
.8..
them by up to 7 Many different combinations of side-chains are found to produce helix
interfaces that are comparably well packed, as if the tertiary structure had been preserved
by a patchwork of improvisations.
X-ray analysis has Shown that despite the great complexity of hemoglobin, the stereo
chemical mechanism of the cooperative effects is comparatively simple. The deoxy or
T -structure is constrained by hydrogen bonds between and within the subunits which
oppose the changes of tertiary structure required for combination with oxygen, while in
the oxy or R-structure these bonds are absent. Some of these bonds are between anionic
groups with pKa'S below 4 and cationic groups with pKa's between 6 and 8. The closure of
these hydrogen bonds leads to a rise and the rupture to a drop in the pKa'S of the cationic
groups. These changes in pKa's are responsible for the Bohr effect. There have been reports
of NMR experiments which purported to disprove this simple me<:hanism, but these have
now been found to have been based on an incorrect assignment of NMR resonances.
Experience as a reviewer of grant applications has taught me that many scientists
detest nothing more than simple explanations for complex phenomena. They would prefer
cooperative oxygen binding to be due to transitions between a multitude of states of
unknown structure rather than two structures capable of being determined by X·ray
analysis; they would like the Bohr effect to arise from a host of small and inexplicable
changes in pK , they would like the hydrogen bonds that you can see to make no signif
a
icant contribution to cooperativity, and those that you cannot see to be responsible for
the cooperative effects.
By contrast, I believe that there is no longer any great mystery about the cooperative
effects of vertebrate hemoglobins, and that the seemingly complex cooperative interactions
in invertebrate hemoglobins and hemocyanins reported in this volume will find similarly
simple explanations once their structure has been determined by X-ray analysis.
May. 1985 M.F. PERUTZ
Contents
Structure and Function of Hemerythrins 1
W.A. Hendrickson, J.L. Smith, S. Sheriff
The Structure of Erythrocruorins and Chlorocruorins, 9
the Invertebrate Extracellular Hemoglobins
S.N. Vinogradov
Physiological Adaptations and Subunit Diversity in Hemocyanins 21
J. Bonaventura, C. Bonaventura
Functions and Functioning of Crustacean Hemocyanin "
B. McMahon
Primary Structure of Arthropod Hemocyanins 59
B. Linzen, W. Schartau, H• • J. Schneider
Quaternary Structure of Arthropod Hemocyanin "
J. Lamy, J. Lamy, P.· Y. Sizaret, P. Billiald, G. Motta
Cephalopod Hemocyanins: Structure and Function 87
K.E. van Holde, K.I. Miller
Molecular and Cellular Adaptations of Fish Hemoglobin-Oxygen 97
Affinity to Environmental Changes
D.A. Powers
Ol(ygen Transport Proteins: A Unitarian View Based on Thermodynamic 125
Kinetic and Stereochemical Consideratioll8
B. Giardina, M. Coletta, L. Zolla, M. Brunori
Evolution and Adaptation of Avian and Crocodilian Hemoglobins 141
A.G. Schriek, C. Paul, J. L~onis
Abnormal Human Hemoglobill8: Molecular Tools 159
to Study Normal Hemoglobin Functions
C. Poysrt, E. Burssux, F. Galac:teros, H. Wacjman
Index 171